外周血中自然杀伤细胞分选纯化方法的比较

Comparison of methods of natural killer cell sorting and purification from human peripheral blood

目的比较荧光激活细胞分选技术（FACS）及免疫磁珠分选技术（MACS）对外周血自然杀伤（NK）细胞分选纯化的效率，为外周血中低比例细胞的深入研究创造基本条件。方法选择20名健康志愿者及3例造血干细胞移植（HSCT）后早期（＋14d）患者，通过FACS及MACS各自分选其外周血中NK细胞。结果分选前健康志愿者外周血NK细胞百分数为（12．86±3．62）％，FACS分选后NK细胞纯度达（96．15±2．03）％，回收率为（95．08±2．16）％；MACS分选后纯度达（93．35±3．61）％，回收率为（94．11±3．01）％。移植后早期患者外周血中NK细胞百分数为（11．01±2．08）％，FACS分选后纯度达（96．22±2．16）％，回收率为（95．27±1．18）％；MACS分选后纯度达（90．98±1．94）％，回收率为（94．54±3．52）％。两种方法分选NK细胞纯度差异有统计学意义（t=5．925，P〈0．05），而回收率差异无统计学意义（t=0．789，P〉0．05）。结论FACS较MACS可以更高效、快速、简便地分离纯化外周血中NK细胞，尤其适于分选移植后早期重建的NK细胞。

Objective To apply fluorescence-activated cell sorting （FACS） and magnetic activated cell sorting （MACS） in sorting natural killer （NK） cells from human peripheral blood. The efficiency of these two methods were compared so that the basic conditions for the further Study on the lower proportion of peripheral blood cells could be created. Methods Twenty eases of normal peripheral blood and 3 cases of post-hematopoietic stem cell transplantation （post-HSCT） were collected and NK cell were sorted by FACS and MACS. Results Before sorting the normal peripheral blood, the proportion of the NK cell in the leukocyte was （12.86±3.62） %, the purity of NK cell was up to （96.15±2.03） % and the recovery was （95.08±2.16） % after sorting by FACS, the purity was up to （93.35±3.61） % and the recovery was （94.11±3.01） % by MACS. And before sorting the post-HSCT peripheral blood, the proportion of the NK cell in the leukocyte was （11.01±2.08） %, the purity of NK cell was up to （96.22±2.16） % and the recovery was （95.27±1.18） % after sorting by FACS. The purity was up to （90.98±1.94） % and the recovery was （94.54±3.52） % by MACS. There is difference in purity of NK cell （t = 5.925, P 〈0.05）, while no difference in rate of recovery between 2 methods （t = 0.789, P 〉0.05）. Conclusion FACS is more efficient, rapidly and easily than MACS in sorting NK cell, especially for post-HSCT cases.